SARS-CoV-2 viral entry and replication is impaired in Cystic Fibrosis airways due to ACE2 downregulation

As an inherited disorder characterized by severe pulmonary disease, cystic fibrosis could be considered a comorbidity for coronavirus disease 2019. Instead, current clinical evidence seems to be heading in the opposite direction. To clarify whether host factors expressed by the Cystic Fibrosis epithelia may influence coronavirus disease 2019 progression, here we describe the expression of SARS-CoV-2 receptors in primary airway epithelial cells. We show that angiotensin converting enzyme 2 (ACE2) expression and localization are regulated by Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) channel. Consistently, our results indicate that dysfunctional CFTR channels alter susceptibility to SARS-CoV-2 infection, resulting in reduced viral entry and replication in Cystic Fibrosis cells. Depending on the pattern of ACE2 expression, the SARS-CoV-2 spike (S) protein induced high levels of Interleukin 6 in healthy donor-derived primary airway epithelial cells, but a very weak response in primary Cystic Fibrosis cells. Collectively, these data support that Cystic Fibrosis condition may be at least partially protecting from SARS-CoV-2 infection.

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The raw data generated in this study are provided in the Source Data file.
Age and gender of patients and healthy donors from whom hNEC and hBEC were isolated are reported in the manuscript, according to data provided by the supplier (Epithelix, Plan-les-Ouates, Switzerland).
Genotype of patients from whom hNEC and hBEC were isolated is reported in the manuscript, according to data provided by the supplier (Epithelix, Plan-les-Ouates, Switzerland).

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Although this study has been approved by the Ethics Committee of the AOUI Verona (Approval No. 2917CESC), in this work all human samples were supplied by Epithelix (Geneva, Switzerland) as commercially available tissues (MucilAir). Thus, we did not utilized any other human sample.
No sample size calculation was performed. A sample size of n=3 is commonly used in biological studies using cell cultures. However, we employed two primary human nasal cells (hNEC) and seven bronchial cells (hBEC) isolated from patients with CF. In addition, we employed a pool of hNEC obtained from 14 subjects and eight primary hBEC samples. All human samples were commercially available from Epithelix (Planles-Ouates, Switzerland). ID of primary hNEC and hBEC with clinical and genetics characteristics have been included in supplementary informations.
No data were excluded from the analysis Experiments were performed in triplicate, indipendedent biological repeats, unless specified in figure legends Since we only used cell culture, randomization was not applicable Since we only used cell culture, blinding was not applicable nature portfolio | reporting summary The commercially obtained cell lines were authenticated by suppliers. CFBE41o-and 16HBE14o-parental cell lines were generated by Gruenert D. (University of California, San Francisco). 16HBE14o-were edited at the endogenous CFTR locus using CRISPR/CAS9 to create isogenic cell lines W1282X. During immortalization of parental cells 16HBE14o-, SV40 sequence was inserted into one of the two CFTR alleles in these cells, rendering them mono-allelic with respect to functional CFTR expression. Full SV40 sequence information and whole genome sequencing data for CFBE41o-and 16HBE14o-parental cell